Enzyme kinetics are the studies of the biophysical properties of enzymes and their substrates.

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Is there a formula for determining the % of ligand bound to a given receptor in a mixture with multiple ligands and receptors?

When a ligand and receptor interact with a given kd, one can easily determine % of ligand bound to the receptor when the kd, total concentration of ligand, and total concentration of receptor are ...
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Enzymatic error rate

I am aware that each enzyme generate a certain amount of misproducts. This is well documented, for example, for the DNA polymerase. I am interested in enzyme involved in biochemical processes, so for ...
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72 views

Enzyme Assay - pectinase

During assaying an enzyme at high temperature, the substrate (Pectin) is degraded by the high temperature rather than by enzyme, so, how can I minimize degradation of the substrate by the temperature? ...
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Does enzyme Km comparisons make sense?

I have encountered comparisons of the Michaelis-menton constant (Km) a few times. They say if Km of an enzyme is higher, then it's affinity to its substrate is lower. How does this make sense? Maybe ...
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Mode of Enzymatic Inhibition via R-Allele

The photo above shows the effects of the R allele of the pea shape gene on the synthesis of an enzyme that converts unbranched starch into branched starch. The r allele of this gene determines an ...
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Kinetic binding / unbinding constants of protein + mRNA ⇌ mRNP

I wish to find an order of magnitude for the binding/unbinding constants of mRNA and RNA-binding proteins. Suppose molecules A and B reversibly associate to give the complex C. Assuming the diffusion ...
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244 views

What's the difference between Ki and IC50?

I have just read this post at researchgate: "How to calculate experimental binding free energy from the IC50 value" I am not be very sure about what the difference is between Ki and IC50. Could ...
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301 views

Why would an Eadie-Hofstee Plot be non-linear? [closed]

Besides cooperativity between multiple active sites on an enzyme, what are the other reasons for the Eadie-Hofstee plot to be non-linear?
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can a bacterial protease inhibitor cocktail be used for Western Blotting involving HUVEC cells and HT-29 adenocarcinoma cells?

Dear fellow biochemists, I need some advice on Western Blotting, more specifically the use of certain protease inhibitors with the RIPA cell lysis buffer and protease inhibitor cocktail. A Millipore ...
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74 views

Pectinase Enzyme Assay

I am working on pectinase enzyme assay. I incubated 900 ul of substrate for 10 minutes in the water bath, followed by adding 2ml of DNSA reagent, then 100ul of enzyme extract added finally i read the ...
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181 views

DpnI over-digestion

We have a long protocol that we are optimizing that includes DpnI digestion of a PCR product (to remove any of the template DNA if it's methylated, and while we're not certain in the blind tests, ...
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What does “thermodynamic equilibrium” mean for an enzyme-substrate complex?

From Fersht, Enzyme Structure and Mechanism p. 87: The Michaelis-Menten mechanism assumes that the enzyme-substrate complex is in thermodynamic equilibrium with free enzyme and substrate. In ...
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Enzyme kinetics [closed]

I can't understand how to study enzyme kinetics. Say I have a lipase and want to study the kinetics of this lipase using a fluorogenic substrate, how would I do this? From what I understand I would ...
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Activity of glucokinase

From Solomon et al, 2013 ACC Synthetic biology and from this video : Here, there are 2 competing reactions for glucose - one with glk as enzyme and other with gdh as enzyme. In the graph, y axis ...
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Are there any methods to quantify H2O2 (hydrogen peroxide) which don't rely on horseradish peroxidase?

I can't use HRP since my substrates interfere with it and I need a real-time method because I want to measure kinetic parameters of some oxidases.
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460 views

Why are enzyme-catalysed reactions slower at lower substrate concentration?

Suppose I'm using 200 nmoles of enzyme and 2 mmoles of substrate. The enzyme should be saturated but if I use 50 mmoles of substrate, the reaction will be faster. Why? I just can't get it! Even at ...
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Modeling yeast biochemical pathway using enzyme kinetics

I need enzyme concentration and metabolites concentration values to en-corporate these values in my model and do some simulation. I searched through some database yeast metabolome database ...
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246 views

What is the use of futile reaction cycling such as Fruc-P to Fruc-BP?

One rate-limiting step of glycolysis is the conversion of Fructose-Phosphate (Fruc-P) to Fructose-Bisphosphate (Fruc-BP), catalysed by Phosphofructokinase (PFK). The reaction involves hydrolysing one ...